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Differentiation of human umbilical cord matrix mesenchymal stem cells into neural-like progenitor cells and maturation into an oligodendroglial-like lineage.

Leite C, Silva NT, Mendes S, Ribeiro A, de Faria JP, Lourenço T, dos Santos F, Andrade PZ, Cardoso CM, Vieira M, Paiva A, da Silva CL, Cabral JM, Relvas JB, Grãos M - PLoS ONE (2014)

Bottom Line: In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples.The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity.The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

View Article: PubMed Central - PubMed

Affiliation: Biocant - Technology Transfer Association, Biocant Park, Cantanhede, Portugal.

ABSTRACT
Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

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Multilineage differentiation potential and immunophenotypic characterization of hUCM-MSCs.The multilineage differentiation potential of hUCM-MSCs was demonstrated after 14 days in culture, under conditions that favour chondrogenic, osteogenic or adipogenic differentiation. Phase contrast images of cells stained with Alcian Blue for chondrogenesis (A), ALP and von Kossa for osteogenesis (B) or Oil Red-O for adipogenesis (C). Scale bar represents 100 µm. For immunophenotypic characterization, hUCM-MSCs at passage 2 were dissociated using Accutase (Life Technologies), labelled with antibodies against the indicated antigens and analysed by flow cytometry. Cells were positive for CD105 (D), CD73 (E), CD90 (F) and CD13 (G) and negative for CD34 (H), NGFR (I), CD11b (J) and CD45 (K) (pink lines) when compared with unlabelled MSCs (green lines), as depicted in the histograms. Histograms were obtained from one sample (UCM#2) at passage 2 and are representative of 3 independent samples at P2 and P8.
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pone-0111059-g002: Multilineage differentiation potential and immunophenotypic characterization of hUCM-MSCs.The multilineage differentiation potential of hUCM-MSCs was demonstrated after 14 days in culture, under conditions that favour chondrogenic, osteogenic or adipogenic differentiation. Phase contrast images of cells stained with Alcian Blue for chondrogenesis (A), ALP and von Kossa for osteogenesis (B) or Oil Red-O for adipogenesis (C). Scale bar represents 100 µm. For immunophenotypic characterization, hUCM-MSCs at passage 2 were dissociated using Accutase (Life Technologies), labelled with antibodies against the indicated antigens and analysed by flow cytometry. Cells were positive for CD105 (D), CD73 (E), CD90 (F) and CD13 (G) and negative for CD34 (H), NGFR (I), CD11b (J) and CD45 (K) (pink lines) when compared with unlabelled MSCs (green lines), as depicted in the histograms. Histograms were obtained from one sample (UCM#2) at passage 2 and are representative of 3 independent samples at P2 and P8.

Mentions: MSCs are multipotent stem cells that can differentiate in vitro into chondrocytes, osteocytes and adipocytes [2]. The multilineage differentiation potential of UCM-MSCs was demonstrated in culture (Figure 2A-C), under conditions that favor osteogenic, adipogenic or chondrogenic differentiation (Materials and Methods section). Chondrogenic induction was observed by Alcian Blue staining (Figure 2A), while osteogenic differentiation was evident by an increase in alkaline phosphatase (ALP) activity (reddish areas) and enhanced mineralization showed by von Kossa staining (dark areas), as shown in Figure 2B. Adipogenic induction, although less efficient, was visible by the cellular accumulation of lipid-rich vacuoles that stained with Oil Red-O (Figure 2C). These results indicate that UCM-MSCs possess the multilineage differentiation capacity characteristic of MSCs [2].


Differentiation of human umbilical cord matrix mesenchymal stem cells into neural-like progenitor cells and maturation into an oligodendroglial-like lineage.

Leite C, Silva NT, Mendes S, Ribeiro A, de Faria JP, Lourenço T, dos Santos F, Andrade PZ, Cardoso CM, Vieira M, Paiva A, da Silva CL, Cabral JM, Relvas JB, Grãos M - PLoS ONE (2014)

Multilineage differentiation potential and immunophenotypic characterization of hUCM-MSCs.The multilineage differentiation potential of hUCM-MSCs was demonstrated after 14 days in culture, under conditions that favour chondrogenic, osteogenic or adipogenic differentiation. Phase contrast images of cells stained with Alcian Blue for chondrogenesis (A), ALP and von Kossa for osteogenesis (B) or Oil Red-O for adipogenesis (C). Scale bar represents 100 µm. For immunophenotypic characterization, hUCM-MSCs at passage 2 were dissociated using Accutase (Life Technologies), labelled with antibodies against the indicated antigens and analysed by flow cytometry. Cells were positive for CD105 (D), CD73 (E), CD90 (F) and CD13 (G) and negative for CD34 (H), NGFR (I), CD11b (J) and CD45 (K) (pink lines) when compared with unlabelled MSCs (green lines), as depicted in the histograms. Histograms were obtained from one sample (UCM#2) at passage 2 and are representative of 3 independent samples at P2 and P8.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214693&req=5

pone-0111059-g002: Multilineage differentiation potential and immunophenotypic characterization of hUCM-MSCs.The multilineage differentiation potential of hUCM-MSCs was demonstrated after 14 days in culture, under conditions that favour chondrogenic, osteogenic or adipogenic differentiation. Phase contrast images of cells stained with Alcian Blue for chondrogenesis (A), ALP and von Kossa for osteogenesis (B) or Oil Red-O for adipogenesis (C). Scale bar represents 100 µm. For immunophenotypic characterization, hUCM-MSCs at passage 2 were dissociated using Accutase (Life Technologies), labelled with antibodies against the indicated antigens and analysed by flow cytometry. Cells were positive for CD105 (D), CD73 (E), CD90 (F) and CD13 (G) and negative for CD34 (H), NGFR (I), CD11b (J) and CD45 (K) (pink lines) when compared with unlabelled MSCs (green lines), as depicted in the histograms. Histograms were obtained from one sample (UCM#2) at passage 2 and are representative of 3 independent samples at P2 and P8.
Mentions: MSCs are multipotent stem cells that can differentiate in vitro into chondrocytes, osteocytes and adipocytes [2]. The multilineage differentiation potential of UCM-MSCs was demonstrated in culture (Figure 2A-C), under conditions that favor osteogenic, adipogenic or chondrogenic differentiation (Materials and Methods section). Chondrogenic induction was observed by Alcian Blue staining (Figure 2A), while osteogenic differentiation was evident by an increase in alkaline phosphatase (ALP) activity (reddish areas) and enhanced mineralization showed by von Kossa staining (dark areas), as shown in Figure 2B. Adipogenic induction, although less efficient, was visible by the cellular accumulation of lipid-rich vacuoles that stained with Oil Red-O (Figure 2C). These results indicate that UCM-MSCs possess the multilineage differentiation capacity characteristic of MSCs [2].

Bottom Line: In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples.The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity.The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

View Article: PubMed Central - PubMed

Affiliation: Biocant - Technology Transfer Association, Biocant Park, Cantanhede, Portugal.

ABSTRACT
Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

Show MeSH
Related in: MedlinePlus